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A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia

Hypoxia is known to stimulate mitochondrial reactive oxygen species (mROS) in cells. Here, we present a detailed protocol to detect mROS using MitoSOX staining in live cells under normoxia and hypoxia. Flow cytometry allows sensitive and reliable quantification of mROS by FlowJo software. We optimiz...

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Detalles Bibliográficos
Autores principales: Yang, Yun, Zhang, Guimin, Yang, Tao, Gan, Jia, Xu, Lin, Yang, Hanshuo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8086139/
https://www.ncbi.nlm.nih.gov/pubmed/33997804
http://dx.doi.org/10.1016/j.xpro.2021.100466
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author Yang, Yun
Zhang, Guimin
Yang, Tao
Gan, Jia
Xu, Lin
Yang, Hanshuo
author_facet Yang, Yun
Zhang, Guimin
Yang, Tao
Gan, Jia
Xu, Lin
Yang, Hanshuo
author_sort Yang, Yun
collection PubMed
description Hypoxia is known to stimulate mitochondrial reactive oxygen species (mROS) in cells. Here, we present a detailed protocol to detect mROS using MitoSOX staining in live cells under normoxia and hypoxia. Flow cytometry allows sensitive and reliable quantification of mROS by FlowJo software. We optimized several aspects of the procedure including hypoxic treatment, working concentrations of the staining buffer, and quantitative analyses. Here, we use HepG2 cells, but the protocol can be applied to other cell lines. For complete details on the use and execution of this protocol, please refer to Yang et al. (2020).
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spelling pubmed-80861392021-05-13 A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia Yang, Yun Zhang, Guimin Yang, Tao Gan, Jia Xu, Lin Yang, Hanshuo STAR Protoc Protocol Hypoxia is known to stimulate mitochondrial reactive oxygen species (mROS) in cells. Here, we present a detailed protocol to detect mROS using MitoSOX staining in live cells under normoxia and hypoxia. Flow cytometry allows sensitive and reliable quantification of mROS by FlowJo software. We optimized several aspects of the procedure including hypoxic treatment, working concentrations of the staining buffer, and quantitative analyses. Here, we use HepG2 cells, but the protocol can be applied to other cell lines. For complete details on the use and execution of this protocol, please refer to Yang et al. (2020). Elsevier 2021-04-20 /pmc/articles/PMC8086139/ /pubmed/33997804 http://dx.doi.org/10.1016/j.xpro.2021.100466 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Yang, Yun
Zhang, Guimin
Yang, Tao
Gan, Jia
Xu, Lin
Yang, Hanshuo
A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia
title A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia
title_full A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia
title_fullStr A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia
title_full_unstemmed A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia
title_short A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia
title_sort flow-cytometry-based protocol for detection of mitochondrial ros production under hypoxia
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8086139/
https://www.ncbi.nlm.nih.gov/pubmed/33997804
http://dx.doi.org/10.1016/j.xpro.2021.100466
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